Utilização da proteína EMA-2 recombinante de Theileria equi expressa em Pichia pastoris, como imunobiológico

Abstract

Theileriosis, endemic disease in Brazil, is a piroplasmosis caused by intra-erythrocytic protozoan, Theileria equi. Causes losses associated with both clinical factors and restriction on the movement of positive animals. Transmission of T. equi occurs mainly during tick feeding which inoculates their infective forms (sporozoites) in horses, which are the vertebrate hosts. The diagnosis and prevention of this disease is needed in endemic and non endemic areas due to the presence of vectors and protozoa. Different ELISAs platforms have been developed using recombinant antigens. EMA-2 is released into the cytoplasm and erythrocyte membrane, suggesting that one of the first antigens recognized by the immune system. The aim of this study was to evaluate the EMA-2 protein of Theileria equi, expressed in Pichia pastoris, as immunobiological. After expression of EMA-2 glycoprotein, an indirect ELISA was developed. The ELISA demonstrated sensitivity of 90.9% and specificity of 83.3% when compared to the commercial kit cELISA. The protein also showed immunogenicity when test the sera of immunized mice with recombinant EMA-2 by immunofluorescence. As the EMA-2 protein antigenic and immunogenic, it may be used diagnostic tests as well as a promising antigen for the development of subunit vaccines.